Brucella abortus recombinant strain and application thereof

A technology of Brucella abortus and recombinant strains, applied in the direction of bacteria, antibacterial drugs, bacterial antigen components, etc., can solve problems such as difficult operation, difficult identification of immunized animals and clinically diseased animals, and huge cost, so as to promote eradication and purifying effect

Inactive Publication Date: 2010-01-20
CHINA AGRI UNIV
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  • Abstract
  • Description
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  • Application Information

AI Technical Summary

Problems solved by technology

For a long time, the global strategy for the prevention and control of brucellosis has been mainly quarantine-culling (in developed countries) or immunization (in developing countries). The former uses quarantine and culling of sick cattle, sheep, pigs and other animals as a measure, which costs a lot of money , Difficult to operate; the latter can prevent the spread of the disease in the animal population through immunization and auxiliary culling measures, but it is difficult to distinguish between immunized animals and clinically diseased animals
Although rough strain vaccines allow for differential diagnosis, allowing the distinction of immunized animals from clinically infected animals, the protective potency of this type of vaccine is significantly lower

Method used

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  • Brucella abortus recombinant strain and application thereof
  • Brucella abortus recombinant strain and application thereof
  • Brucella abortus recombinant strain and application thereof

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0039] Embodiment 1, the construction of inactivated glycosyltransferase SLT subunit protein gene recombinant strain

[0040] According to the glycosyltransferase SLT subunit protein gene (GenBankAccession Number: BABI_0664) in the Brucella abortus genome, three pairs of primers were designed, one pair of primers was used to amplify the homology arm DA16-1, and the second pair of primers was used to amplify the homology arm DA16-1. DA16-2 was amplified, and the third pair of primers amplified the kan fragment. Homologous arm DA6-1 and homologous arm DA6-2 were connected to form sequence DA6, and sequence DA16 was inserted into pEASY-T1 Simple vector to obtain recombinant vector pEASY-T1-DA16, then digested and inserted into kan resistance gene to obtain recombinant vector pEASY -T1-KDA16. The recombinant vector pEASY-T1-KDA16 was digested to obtain the fragment KDA16, which was inserted into the pBluescripts IIKS vector to obtain the final recombinant vector pBluescripts II K...

Embodiment 2

[0093] Embodiment 2, the application of recombinant bacterial strain NBA16

[0094] Experimental animals: 4-6 weeks old SPF grade Balb / C female mice, purchased from Beijing Weitong Lihua Experimental Animal Technology Co., Ltd., license number SCXK (Beijing) 2009-0001.

[0095] 1. Toxicity identification

[0096] The mice were randomly divided into three groups, 20 in each group. The specific vaccination plan is as follows:

[0097] The first group (test group): inoculate the recombinant bacterial strain NBA16 that embodiment 1 prepares, every mouse intraperitoneally inoculates dose 0.1ml, contains 10 7 CFU (CFU: colony formine unit, colony forming unit, hereinafter referred to as CFU) bacteria.

[0098] The second group (control group 1): inoculate Brucella abortus bacterial strain 2308, every mouse intraperitoneally inoculates dose 0.1ml, contains 10 7 CFU bacteria.

[0099] The third group (control group 2): inoculate Brucella abortus bacterial strain S19, every mouse ...

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Abstract

The invention discloses a brucella abortus recombinant strain and the application thereof, and the brucella abortus is obtained by inactivating glycosyltransferase SLT subunit protein gene in strain 2308. Compared with the original strain, the strain obtained in the invention has little strain toxicity, good immunity protecting effect and potential differential diagnosis mark, and can be used for research and development of attenuated vaccine. The invention is beneficial to the reformation of brucellosis vaccine, the research and the preparation of differential diagnosis reagent and the development of the brucellosis vaccine and the differential diagnosis reagent matched with the vaccine, and has very important significance on promoting the elimination of brucellosis of the animals all over the world.

Description

technical field [0001] The invention relates to a recombinant bacterium of Brucella abortus and its application, in particular to a recombinant bacterium of an inactivated glycosyltransferase SLT subunit protein gene and its application. Background technique [0002] Brucellosis is a zoonotic infectious disease, second only to foot-and-mouth disease among large animal infectious diseases, and has important political and economic influences. The epidemic situation in countries around the world is very serious. For a long time, the global strategy for the prevention and control of brucellosis has been mainly quarantine-culling (in developed countries) or immunization (in developing countries). The former uses quarantine and culling of sick cattle, sheep, pigs and other animals as a measure, which costs a lot of money , Difficult to operate; the latter can prevent the spread of the disease in the animal population through immunization and auxiliary culling measures, but it is d...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): C12N1/21A61K39/10A61P31/04C12R1/01
Inventor 吴清民王真任婕张春燕牛建蕊董浩崔波刘文娟刘文晓
Owner CHINA AGRI UNIV
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